本文選題：乳腺癌 + 外周血��； 參考：《電子科技大學(xué)》2016年博士論文

【摘要】：乳腺癌是常見的女性惡性腫瘤,具有異質(zhì)性高的特點(diǎn),即使細(xì)胞形態(tài)學(xué)和雌激素受體等細(xì)胞分子表型很相似的乳腺癌患者,也可能具有不同的預(yù)后表現(xiàn),或?qū)ο嗤闹委煼桨复嬖诓煌姆磻?yīng)。因此,除目前臨床常用的指標(biāo)外,需要更準(zhǔn)確、更具臨床實(shí)用性的預(yù)測(cè)因子,為乳腺癌個(gè)體化治療提供依據(jù)。腫瘤的發(fā)展不僅由腫瘤細(xì)胞自身決定,也依賴于腫瘤細(xì)胞與間質(zhì)之間復(fù)雜的相互作用。作為腫瘤免疫效應(yīng)的執(zhí)行場(chǎng)所,腫瘤微環(huán)境中包含了大量的免疫細(xì)胞,為腫瘤免疫逃逸提供條件,乳腺癌的免疫微環(huán)境是影響其發(fā)展和預(yù)后的重要因素之一。外周血是用于疾病診斷的最常用材料,被患者廣泛接受,是無創(chuàng)性分子診斷的理想材料。而且,相比于腫瘤組織于手術(shù)中一次取樣,忽略了微環(huán)境動(dòng)態(tài)變化的特點(diǎn),外周血可定期多次取樣,便于監(jiān)測(cè)。在乳腺癌發(fā)展的過程中,作為腫瘤微環(huán)境中免疫細(xì)胞的主要來源,關(guān)于乳腺癌患者外周血與腫瘤局部免疫微環(huán)境的關(guān)系,外周血是否能夠反映腫瘤局部免疫微環(huán)境的變化,以及外周血來源的免疫微環(huán)境相關(guān)分子,目前尚無深入、系統(tǒng)的研究。腫瘤生物信息學(xué)是一種新興的手段,將生物信息學(xué)的方法學(xué)、工具軟件及數(shù)據(jù)庫等應(yīng)用于腫瘤的分子機(jī)制、新的腫瘤生物標(biāo)志物和個(gè)體化治療措施的研究中。本研究以基因表達(dá)譜數(shù)據(jù)為基礎(chǔ),聯(lián)合生物信息學(xué)工具及文獻(xiàn)挖掘手段,首先分析了良性和惡性乳腺腫瘤外周血單個(gè)核細(xì)胞的基因表達(dá)變化,評(píng)價(jià)外周血作為替代材料,用于評(píng)估乳腺癌患者的免疫狀態(tài)可能性。然后,比較了乳腺癌外周血細(xì)胞與癌間質(zhì)細(xì)胞差異基因表達(dá)的一致性,評(píng)價(jià)外周血與癌間質(zhì)免疫微環(huán)境的關(guān)系。最后,篩選了乳腺癌組織與外周血細(xì)胞之間的交互作用分子,探討乳腺癌組織對(duì)外周免疫系統(tǒng)可能的遠(yuǎn)程調(diào)節(jié)。本研究主要內(nèi)容包括如下三部分:1、良性與惡性乳腺腫瘤外周血細(xì)胞基因差異表達(dá)研究。從GEO數(shù)據(jù)庫中獲取良性和惡性乳腺腫瘤患者外周血單個(gè)核細(xì)胞(PBMCs)mRNA表達(dá)譜。包括57例乳腺癌,37例良性乳腺腫瘤和31例健康人外周血標(biāo)本。GEO2R工具分別篩選良性和惡性乳腺腫瘤相較于健康人群的外周血差異表達(dá)基因,DAVID工具富集基因功能和通路。STRING數(shù)據(jù)庫構(gòu)建差異表達(dá)基因蛋白產(chǎn)物相互作用的網(wǎng)絡(luò),篩選核心基因。良性和惡性乳腺腫瘤分別篩選到563和237個(gè)差異基因,兩類患者外周血基因表達(dá)模式存在顯著差異,其中乳腺癌差異基因功能側(cè)重于免疫反應(yīng),涉及白細(xì)胞的激活、血管生成等生物學(xué)過程以及白細(xì)胞跨內(nèi)皮遷移信號(hào)通路。IL-8、rhob、itgb1等為關(guān)鍵基因,可能在乳腺癌外周免疫激活中起關(guān)鍵作用。2、乳腺癌患者外周血與腫瘤間質(zhì)免疫微環(huán)境關(guān)系研究。下載geo數(shù)據(jù)庫中乳腺癌間質(zhì)及外周血細(xì)胞mrna表達(dá)譜。乳腺癌間質(zhì)樣本來源于53例乳腺癌組織,31例匹配的癌旁正常組織。brb-arraytools預(yù)處理數(shù)據(jù)并篩選差異表達(dá)基因,david工具富集分析基因功能和通路。兩組數(shù)據(jù)相同的差異表達(dá)基因僅為103個(gè),占二者差異基因總數(shù)的2%。乳腺癌間質(zhì)與外周血較正常對(duì)照的差異表達(dá)基因交疊數(shù)較少,但二者差異基因涉及的一致的生物學(xué)功能主要集中在免疫反應(yīng)相關(guān)過程。進(jìn)一步對(duì)差異表達(dá)的細(xì)胞因子的研究發(fā)現(xiàn),pbmcs表達(dá)譜差異表達(dá)基因中共81個(gè)屬于細(xì)胞因子、趨化因子及其互作基因(其中76個(gè)表達(dá)上調(diào),5個(gè)下調(diào))。乳腺癌間質(zhì)表達(dá)譜中共103個(gè)屬于細(xì)胞因子、趨化因子及其互作基因(其中55個(gè)表達(dá)上調(diào),48個(gè)下調(diào))。乳腺癌外周血細(xì)胞與間質(zhì)的細(xì)胞因子相關(guān)基因的表達(dá)模式具有相似之處,其共同擾動(dòng)的通路,均表現(xiàn)為免疫抑制和促腫瘤效應(yīng)。3、乳腺癌組織與外周血的交互作用研究。下載geo數(shù)據(jù)庫中乳腺癌間質(zhì)、外周血細(xì)胞及乳腺癌上皮細(xì)胞mrna表達(dá)譜。乳腺癌上皮細(xì)胞樣本來源于28例浸潤性乳腺癌組織,5例正常乳腺組織。brb-arraytools預(yù)處理數(shù)據(jù)并篩選差異表達(dá)基因。利用uniprot、string、dlrp蛋白質(zhì)相互作用數(shù)據(jù)庫,對(duì)乳腺癌上皮細(xì)胞(間質(zhì)細(xì)胞)差異表達(dá)上調(diào)的基因蛋白產(chǎn)物進(jìn)行亞細(xì)胞定位,確定差異表達(dá)的分泌蛋白,及與其相互作用的乳腺癌pbmcs膜蛋白,作為分子對(duì)話中的受體。檢索與受體相互作用的胞內(nèi)下游蛋白質(zhì);使用cytoscape軟件構(gòu)建配體—受體—胞內(nèi)下游蛋白質(zhì)的相互作用網(wǎng)絡(luò)。david工具對(duì)互作分子進(jìn)行基因功能和通路富集。乳腺癌上皮細(xì)胞與pbmcs之間共得到101對(duì)相互作用分子,其中fn1、itgb1、flt1、comp、cxcl12為關(guān)鍵蛋白。乳腺癌間質(zhì)細(xì)胞與pbmcs之間得到包括19個(gè)表達(dá)上調(diào)的乳腺癌間質(zhì)細(xì)胞分泌蛋白(配體),26個(gè)pbmcs膜(受體),73個(gè)與pbmcs受體互作的胞內(nèi)蛋白的交互作用網(wǎng)絡(luò),其中hub基因?yàn)閕tgb1,itgb3,itga2b,cxcr4。交互作用網(wǎng)絡(luò)中的基因功能均主要涉及細(xì)胞粘附、細(xì)胞遷移、白細(xì)胞激活、免疫反應(yīng)等生物學(xué)過程,參與的信號(hào)通路主要有細(xì)胞外基質(zhì)—受體相互作用,細(xì)胞因子—細(xì)胞因子受體相互作用,趨化因子信號(hào)通路等。綜上所述,利用基因表達(dá)譜芯片數(shù)據(jù)及生物信息學(xué)工具和方法,本論文探討了乳腺癌患者外周血細(xì)胞的基因差異表達(dá),分析了外周血與乳腺癌間質(zhì)微環(huán)境在免疫反應(yīng)方面的相似性。通過乳腺癌組織與外周血細(xì)胞可能的相互作用分子網(wǎng)絡(luò)的分析,探討這種相似性的分子機(jī)制。本研究發(fā)現(xiàn),il-8、rhob、itgb1、fn1、flt1、comp、cxcl12、itgb3、itga2b、cxcr4等基因可能在乳腺癌患者以外周血為基礎(chǔ)的免疫微環(huán)境的形成中發(fā)揮著重要作用。其中itgb1和cxcr4在乳腺癌外周血細(xì)胞顯著上調(diào),同時(shí)為乳腺癌細(xì)胞和間質(zhì)細(xì)胞分泌蛋白共同的作用靶點(diǎn),對(duì)其深入研究,干預(yù)腫瘤來源的分子與外周血細(xì)胞的遠(yuǎn)程相互作用,有可能改變外周免疫細(xì)胞向腫瘤局部的趨化,為乳腺癌的免疫治療及預(yù)后監(jiān)測(cè)提供新的思路。
[Abstract]:Breast cancer is a common female malignant tumor with high heterogeneity, and breast cancer patients with similar cell morphology and estrogen receptor phenotype are likely to have different prognosis or different responses to the same treatment. The more clinically practical predictors provide a basis for the individualized treatment of breast cancer. The development of the tumor is not only determined by the tumor cells themselves, but also depends on the complex interaction between the tumor cells and the interstitium. As the execution place of the tumor immune effect, a large number of immune cells are contained in the tumor microenvironment and escape from the tumor immunity. The immune microenvironment of breast cancer is one of the most important factors affecting the development and prognosis of breast cancer. Peripheral blood is the most commonly used material for diagnosis of disease. It is widely accepted by patients and is an ideal material for noninvasive molecular diagnosis. Moreover, compared to the one sampling in the operation of the tumor tissue, the characteristics of the dynamic changes in the microenvironment are ignored. Blood can be sampled regularly to facilitate monitoring. In the development of breast cancer, as the main source of immune cells in the tumor microenvironment, the relationship between peripheral blood and local immune microenvironment of the breast cancer patients, whether the peripheral blood can reflect the changes in the local immune microenvironment of the tumor, and the immune microenvironment of the peripheral blood There is no deep and systematic research on the biomarkers. Tumor bioinformatics is a new means to apply bioinformatics methodology, tool software and database to the molecular mechanism of tumor, new tumor biomarkers and individualized treatment. Informatics tools and literature mining methods first analyzed the gene expression changes in peripheral blood mononuclear cells of benign and malignant breast tumors, and evaluated the peripheral blood as an alternative material to evaluate the immune status of breast cancer patients. Then, the conformance of differentially expressed genes between the peripheral blood cells of breast cancer and the interstitial cells of breast cancer was compared. To evaluate the relationship between peripheral blood and intercellular immune microenvironment. Finally, the interaction molecules between breast cancer tissue and peripheral blood cells were screened and the possible remote regulation of the peripheral immune system of breast cancer tissues was explored. The main contents of this study include the following three parts: 1, the differential expression of the peripheral blood cell genes in benign and malignant breast tumors The expression profiles of peripheral blood mononuclear cells (PBMCs) mRNA in benign and malignant breast tumors were obtained from the GEO database, including 57 cases of breast cancer, 37 benign breast tumors and 31 healthy human peripheral blood samples by.GEO2R tool, respectively, to screen the peripheral blood differential expression genes of benign and malignant breast tumors compared to the Yu Jian Kang population, DAVID tool The gene function and pathway.STRING database constructed the network that differentially expressed gene protein products and screened the core genes. 563 and 237 differential genes were screened for benign and malignant breast tumors. There were significant differences in the expression patterns of peripheral blood gene in the two types of patients. The differential gene function of breast cancer focused on the immune response. The biological processes, such as leukocyte activation, angiogenesis and other biological processes, such as.IL-8, RhoB, itgb1, and so on, may play a key role in the peripheral immune activation of breast cancer, and the relationship between peripheral blood and tumor interstitial immune microenvironment in breast cancer patients. Downloading the interstitial and external breast cancer in the geo database. MRNA expression profiles of peripheral blood cells. The samples of breast cancer were derived from 53 breast cancer tissues, 31 matched paracancerous normal tissues with.Brb-arraytools preprocessing data and screening differentially expressed genes. The David tool enriched the function and pathway of the gene. The same differentially expressed genes in the two groups were only 103, accounting for the total number of differences in the total number of two 2%.. The differential expression of differentially expressed genes between the interstitial and peripheral blood of the breast cancer was less than that of the normal control, but the consistent biological function involved in the two differentially expressed genes was mainly concentrated in the immune response related process. Further research on the differentially expressed cytokines showed that the difference table of PBMCs expression spectrum reached 81 cytokine, and the chemokines were chemotactic. Gene expression profiles (76 up-regulated and 5 down-regulated). The expression profiles of mammary carcinoma were 103 cytokine, chemokines and their interaction genes (55 up and 48 down-regulation). The expression patterns of the peripheral blood cells and the cytokine related genes in breast cancer were similar, and their common disturbance pathway The interaction of immunosuppressive and tumor promoting effect.3 and the interaction of breast cancer tissue with peripheral blood. The mRNA expression profiles of breast cancer stroma, peripheral blood cells and breast cancer epithelial cells were downloaded in the geo database. The breast cancer epithelial cells were derived from 28 cases of invasive breast cancer and 5 normal breast tissues by.Brb-arraytools preconditioning Data and screening differential expression genes. Using UniProt, string, and DLRP protein interaction database, subcellular localization of gene protein products up regulated by differential expression of breast cancer epithelial cells (stromal cells), differentially expressed secretory proteins, and PBMCs membrane protein of breast cancer that are used for their interaction are used as the receptor in the molecular dialogue. To retrieve the intracellular downstream proteins interacting with the receptor, and use the Cytoscape software to construct the ligand receptor - intracellular downstream protein interaction network.David tool for gene function and pathway enrichment. There are 101 pairs of interacting molecules between breast cancer epithelial cells and PBMCs, of which FN1, itgb1, Flt1, comp, CXCL12 is a key protein. Between mammary cancer stromal cells and PBMCs, the intercellular protein (ligand), 26 PBMCs membranes (receptors), and 73 intercellular proteins interacting with PBMCs receptors, are obtained between mammary carcinoma cells and 19 up regulated breast cancer cells, in which the gene function of the hub gene is itgb1, ITGB3, itga2b, and cxcr4. interaction network It is mainly involved in biological processes such as cell adhesion, cell migration, leukocyte activation, and immune response. The signal pathways involved mainly include extracellular matrix receptor interaction, cytokine receptor interaction, chemokine signaling pathway and so on. In summary, the gene expression chip data and bioinformatics tools are used in summary. Methods, the gene differential expression of peripheral blood cells in breast cancer patients was discussed. The similarity between the peripheral blood and breast cancer microenvironment in the immune response was analyzed. The molecular mechanism of the possible interaction of breast cancer tissues and peripheral blood cells was analyzed to explore the molecular mechanism of this similarity. This study found that IL-8, Rho B, itgb1, FN1, Flt1, comp, CXCL12, ITGB3, itga2b, CXCR4 may play an important role in the formation of immune microenvironment based on the peripheral blood of breast cancer patients. Itgb1 and CXCR4 are up up in the peripheral blood cells of breast cancer, at the same time, for the common target of breast cancer cells and interstitial cells to secrete proteins. The long-range interaction between tumor sources and peripheral blood cells may change the chemotaxis of the peripheral immune cells to the tumor, and provide new ideas for the immunotherapy and prognosis monitoring of breast cancer.

【學(xué)位授予單位】：電子科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R737.9

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 欒德琴;白云峰;包文斌;陳國宏;;基因本體論研究進(jìn)展及在生物信息數(shù)據(jù)庫中的應(yīng)用[J];江蘇農(nóng)業(yè)科學(xué);2009年04期

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本文編號(hào)：1789337